LOCUS       sequence-135046-        6991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the PHLS gene as a fusion with the CpcB gene followed by 
            the CmR gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  AUTHORS   Formighieri C, Melis A
  TITLE     A phycocyanin.phellandrene synthase fusion enhances recombinant 
            protein expression and beta-phellandrene (monoterpene) hydrocarbons 
            production in Synechocystis (cyanobacteria).
  JOURNAL   Metab Eng. 2015 Nov;32:116-24. doi: 10.1016/j.ymben.2015.09.010. 
            Epub 2015 Sep 26.
  PUBMED    26410450
REFERENCE   2  (bases 1 to 6991)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab 
            Eng."; date: "2015-11"; volume: "32:116-24. doi"; pages: " 
            10.1016/j.ymben.2015.09.010. Epub 2015 Sep 26"
COMMENT     Sequence Label: CpcB•PHLS+Cpc
FEATURES             Location/Qualifiers
     source          1..6991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3382..3484
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3485..4144
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3551..3570)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4806..4824)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4845..4861)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4845..4861)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4858..4880)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4869..4885
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4893..4899,4900..4917,4918..4923))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4938..4959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5076..5093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5247..5835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5327..5346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6006..6797,6798..6866))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6629..6648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6867..6971)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gagaagagtc cctgaatatc aaaatggtgg gataaaaagc tcaaaaagga
      721 aagtaggctg tggttcccta ggcaacagtc ttccctaccc cactggaaac taaaaaaacg
      781 agaaaagttc gcaccgaaca tcaattgcat aattttagcc ctaaaacata agctgaacga
      841 aactggttgt cttcccttcc caatccagga caatctgaga atcccctgca acattactta
      901 acaaaaaagc aggaataaaa ttaacaagat gtaacagaca taagtcccat caccgttgta
      961 taaagttaac tgtgggattg caaaagcatt caagcctagg cgctgagctg tttgagcatc
     1021 ccggtggccc ttgtcgctgc ctccgtgttt ctccctggat ttatttaggt aatatctctc
     1081 ataaatcccc gggtagttaa cgaaagttaa tggagatcag taacaataac tctagggtca
     1141 ttactttgga ctccctcagt ttatccgggg gaattgtgtt taagaaaatc ccaactcata
     1201 aagtcaagta ggagattaat tcaatgttcg acgtattcac tcgggttgtt tcccaagctg
     1261 atgctcgcgg cgagtacctc tctggttctc agttagatgc tttgagcgct accgttgctg
     1321 aaggcaacaa acggattgat tctgttaacc gcatcaccgg taatgcttcc gctatcgttt
     1381 ccaacgctgc tcgtgctttg ttcgccgaac agccccaatt aatccaaccc ggtggaaacg
     1441 cctacaccag ccgtcgtatg gctgcttgtt tgcgtgacat ggaaatcatc ctccgctatg
     1501 ttacctacgc aaccttcacc ggcgacgctt ccgttctaga agatcgttgc ttgaacggtc
     1561 tccgtgaaac ctacgttgcc ctgggtgttc ccggtgcttc cgtagctgct ggcgttcaaa
     1621 aaatgaaaga agctgccctg gacatcgtta acgatcccaa tggcatcacc cgtggtgatt
     1681 gcagtgctat cgttgctgaa atcgctggtt acttcgaccg cgccgctgct gccgtagccc
     1741 atatgtgtag tttgcaagtt tctgatccta ttcctaccgg acgccgttcc ggtggttatc
     1801 ccccggcctt atgggatttc gatactattc aatccctgaa taccgaatat aagggcgaac
     1861 gtcacatgcg tcgggaagaa gacttaattg gtcaagttcg ggaaatgttg gtgcacgaag
     1921 tagaagatcc cactccccag ttggaattca ttgacgatct gcataaattg ggcatttcct
     1981 gccattttga aaacgagatt ctgcaaattc tcaaatccat ttatctcaac caaaactata
     2041 aacgggacct ctattctacc agtttagcct tccgtctctt gcgtcaatac gggtttatct
     2101 tgccgcagga agtttttgac tgctttaaaa acgaagaagg tacggatttt aaacccagct
     2161 tcggccggga tattaagggt ctgttacagt tgtacgaagc ctcctttttg tcccggaagg
     2221 gggaagaaac tttacaactc gcccgcgaat ttgctaccaa aatcttgcaa aaggaagtcg
     2281 atgaacggga atttgctact aaaatggaat ttcccagtca ctggaccgta caaatgccta
     2341 acgctcggcc ttttatcgat gcctatcgtc ggcgtcccga catgaacccc gtggttctgg
     2401 aactcgccat tctcgatacc aatatcgtgc aagctcagtt tcaagaagaa ttgaaggaga
     2461 cctcccgttg gtgggaaagc acggggattg ttcaagaact gccgtttgtt cgggaccgga
     2521 ttgtggaagg ttatttttgg accattggtg ttactcaacg ccgtgaacac ggttacgaac
     2581 gtattatgac ggccaaagtc atcgctttgg tgacctgttt ggatgatatt tatgacgtat
     2641 atggcactat tgaagaattg caactcttca cctctacgat tcagcgttgg gatttggagt
     2701 ctatgaagca gttaccgact tatatgcagg taagcttcct ggccttgcac aattttgtaa
     2761 ccgaagtggc ctatgatacg ctgaagaaaa agggctacaa ctctaccccc tatttgcgga
     2821 agacttgggt ggatttggtc gaaagttaca ttaaggaagc cacttggtac tataatgggt
     2881 acaaaccctc tatgcaggaa tacctcaaca acgcctggat ctctgtgggc agcatggcta
     2941 ttttgaatca tttgtttttt cgctttacta atgaacgcat gcataagtac cgggacatga
     3001 atcgtgtatc ctctaatatt gtgcggttag ccgacgatat gggaacctct ttggccgaag
     3061 ttgaacgcgg tgacgtgccc aaagctatcc aatgttacat gaatgaaacg aacgcctctg
     3121 aggaggaggc ccgcgaatat gtgcggcgcg ttatccagga agaatgggaa aaactgaaca
     3181 ctgaactgat gcgcgacgac gacgatgacg atgatttcac cttaagtaaa tactactgcg
     3241 aagtcgttgc taacctgacc cggatggctc agttcattta ccaagatggt tccgatgggt
     3301 ttgggatgaa agattccaaa gtaaatcgtt tactgaaaga aacgctgatt gagcgctatg
     3361 agtgaagatc tgcggccgcg ttgatcggca cgtaagaggt tccaactttc accataatga
     3421 aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc
     3481 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     3541 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     3601 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     3661 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     3721 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     3781 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     3841 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     3901 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     3961 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     4021 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgtct gtgatggctt
     4081 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     4141 gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggggatcctc tggttatttt
     4201 aaaaaccaac tttactcagg ttccataccc gagaaaatcc agcttaaagc tgacatatct
     4261 aggaaaattt tcacattcta acgggagata ccagaacaat gaaaacccct ttaactgaag
     4321 ccgtttccac cgctgactct caaggtcgct ttctgagcag caccgaattg caaattgctt
     4381 tcggtcgtct acgtcaagct aatgctggtt tgcaagccgc taaagctctg accgacaatg
     4441 cccagagctt ggtaaatggt gctgcccaag ccgtttataa caaattcccc tacaccaccc
     4501 aaacccaagg caacaacttt gctgcggatc aacggggtaa agacaagtgt gcccgggaca
     4561 tcggctacta cctccgcatc gttacctact gcttagttgc tggtggtacc ggtcctttgg
     4621 atgagtactt gatcgccggt attgatgaaa tcaaccgcac ctttgacctc tcccccagct
     4681 ggtatgttga agctctgaaa tacatcaaag ctaaccacgg cttgagtggc gatgcccgtg
     4741 acgaagctaa ttcctacctc gattacgcca tcaatgctct gagctaggag ctccagcttt
     4801 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
     4861 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4921 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4981 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5041 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5101 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5161 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5221 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5281 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5341 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5401 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5461 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5521 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5581 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5641 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5701 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5761 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5821 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5881 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5941 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6001 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6061 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6121 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6181 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6241 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6301 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6361 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6421 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6481 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6541 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6601 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6661 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6721 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6781 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6841 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6901 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6961 ggggttccgc gcacatttcc ccgaaaagtg c
//
